Inhibition of human corneal epithelial production of fibrotic mediator TGF-β2 by basement membrane-like extracellular matrix
Inhibition of human corneal epithelial production of fibrotic mediator TGF-β2 by basement membrane-like extracellular matrix
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DOI：
10.1167/iovs.06-0772
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发表时间：
2007-03-01
影响因子：
4.4
通讯作者：
Fini, M. Elizabeth
中科院分区：
文献类型：
--
作者：
LaGier, Adriana J.;Yoo, Sonia H.;Fini, M. Elizabeth
目的.转化生长因子（TGF）-β 2是小鼠角膜修复过程中纤维化标志物表达的主要上皮介质。通过在基底膜样细胞外基质提取物（Matrigel; BD Biosciences，贝德福德，MA）上铺板，减少培养的兔角膜上皮细胞产生TGF-β 2蛋白。本研究的目的是进一步了解基质胶调节的性质。TGF-β 2蛋白、mRNA和基因转录促进在培养的人角膜上皮细胞中进行了表征。基质胶在mRNA积累和基因转录启动子活性水平上抑制TGF-β 2的产生。Matrigel的这种作用不能用（1）生长因子污染物来解释，因为生长因子减少的Matrigel也抑制TGF-β 2;（2）独立的基质组分，因为主要ECM组分层粘连蛋白和胶原IV的纯形式不能再现该作用;或（3）抑制组成型TGF-β 2自分泌反馈环，因为加入外源性TGF-β 2增加p-Smad 3并恢复TGF-β 2 mRNA水平。此外，Matrigel减少TGF-β 2的能力不能用其几何形状来解释，因为将细胞接种在具有类似于Matrigel的三维形貌的合成的生物基质上不会抑制TGF-β 2的产生。基质胶引起ezrin减少，ezrin是ezrin-radixinmoesin（ERM）家族的成员，其通过Rho信号通路在组织中上皮细胞的极性建立中起作用。这些发现表明Matrigel抑制TGF-β 2基因表达，并指出依赖于Matrigel组成和结构的机制。Mattigel减少ezrin的能力与这一想法一致，并将未来研究的重点指向ERM/Rho途径。
PURPOSE. Transforming growth factor (TGF)-beta 2 is a major epithelial mediator of fibrotic marker expression during corneal repair in mice. Production of TGF-beta 2 protein by cultured rabbit corneal epithetial cells is reduced by plating on a basement membrane-like extracellular matrix extract (Matrigel; BD Biosciences, Bedford, MA). The goal of the present study was to understand further the nature of Matrigel regulation.METHODS. TGF-beta 2 protein, mRNA, and gene transcriptional promotion were characterized in cultured human corneal epithelial cells.RESULTS. TGF-beta 2 production was inhibited by Matrigel at the level of mRNA accumulation and activity of the gene transcriptional promoter. This effect of Matrigel was not explained by (1) growth factor contaminants, as growth-factor reduced Matrigel also inhibited TGF-beta 2; (2) independent matrix components, as the pure forms of the major ECM components laminin and Collagen IV did not reproduce the effect; or (3) inhibition of a constitutive TGF-beta 2 autocrine feedback loop, as addition of exogenous TGF-beta 2 increased p-Smad3 and restored TGF-beta 2 mRNA levels. In addition, Matrigel's ability to reduce TGF-beta 2 was not explained by its geometry, as TGF-beta 2 production was not inhibited by plating cells on a synthetic nanofiber matrix with a three-dimensional topography similar to Matrigel. Matrigel caused a reduction of ezrin, a member of the ezrin-radixinmoesin (ERM) family, which plays a role in establishing polarity of epithelial cells in tissues through the Rho signaling pathway.CONCLUSIONS. These findings indicate that Matrigel inhibits TGF-beta 2 gene expression and point to a mechanism dependent on Matrigel composition and structure. The capacity of Mattigel to reduce ezrin is consistent with this idea and directs the focus of future studies toward the ERM/Rho pathway.